HumanandMouseReactiveT-betMonoclonalAntibodiesBDBiosciencesnowoerstwoclones,O4-46and4B10,orthestudyotheTh1signaturetranscriptionactor,T-bet,inhumanandmousesamples.T-betisexpressedinresponsetoantigentriggeredTcellreceptor(TCR)signalingandseveralcytokines.T-betexpressionleadstomanycellularresponsesincludingtheexpressionointereron-γ(IFN-γ).T-bet(clones04-46and4B10)puriedandfuorochrome-conjugatedantibodiesThenewBDPharmingen™T-betmonoclonalantibodies(O4-46and4B10)reactwithhumanandmouseT-bet(T-boxgeneexpressedinTcells),amasterregulatorytranscriptionactorthatisalsoknownasTBX21(T-box21)andTBLYM(T-boxtranscriptionactor,expressedinlymphocytes).HumanandmouseT-betproteinsequencesare88%homologous.1AvailableconjugatesincludeAlexaFluor®488,BDHorizonV450,andPerCP-Cy5.5(cloneO4-46only),AlexaFluor®647,andPEormatstoenablemaximumexibilityordesignomulticoloruorescentantibodystainingpanelsincombinationwithanyBDFACS™brandowcytometer.T-betwasdiscoveredthroughayeastone-hybridscreenusingtheTh1-specifcportionotheIL-2promoterlinkedtoareportergene.T-betwasthentransormedwithconstructsromanactivatedTh1cellcDNAlibraryusedtoanactivationdomain.Aterurtherselection,eightcloneswereidentifedthatcontainedsequencesthatwererelatedtooridenticaltoT-bet.1ExpressionoT-betleadstotheexpressionoIFN-γthroughbindingtoenhancersonthepromoterotheIfnγgene.T-beturtherstabilizestheTh1phenotypethroughbindingtoasilencerontheil4gene,preventingtheexpressionotheTh2signaturecytokineIL-4.2InadditiontoTh1cells,T-betisexpressedbyandactivatestranscriptionalactivitieswithinotherhematopoieticcells,includingstemcells,NK,andNKTcellsandsubsetsothymocytes,primed/activatedCD4+Tcells,CD8+Tcells,andγδTcells,Bcells,anddendriticcells.FeaturesUseulmarkerorthedetectionoTh1,NK,andotherIFN-γexpressingcells.Boththe4B10andhigh-sensitivitycloneO4-46reactwithhumanandmouseT-bet.Suitableoruseinseveralapplicationsincludingowcytometry,Westernblot,andimmunohistochemistry.Availableinseveralormatsincludingpurifed,PE,andAlexaFluor®647.High-sensitivitycloneO4-46isalsoavailableinAlexaFluor®488,BDHorizon™V450,andPerCP-Cy™5.5.CompatiblewithBDPhosow™buersystemsincludingBDPhosowLyse/FixBuerandBDPhosowPermeabilizationBuersIthroughIV.
ForResearchUseOnly.Notoruseindiagnosticortherapeuticprocedures.Visitbdbiosciences.com/tcellormoreinormation.Figure1.WholebloodwastreatedwithBDPhoslowLyse/FixBuer(Cat.No.558049).CellswerepermeabilizedwithBDPhoslowPermBuerIII(Cat.No.558050),washed,andstainedwithPEMouseanti–T-betCloneO4-46(Cat.No.561268)orClone4B10(Cat.No.561265),FITCMouseAnti-HumanCD45RO(Cat.No.555492),APCMouseAnti-HumanCD45RA(Cat.No.550855),BDHorizon™V450MouseAnti-HumanCD3(Cat.No.560365),andPerCPMouseAnti-HumanCD4(Cat.No.347324)antibodies.ContourplotsshowingCD45RO(topplot)orCD45RA(bottomplot)versusT-betexpressionpatternsorCD3+CD4+lymphocytesweregeneratedusingaBDFACSCanto™IIlowcytometer.
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